Fluorescent homogeneous immunosensors for detecting pathogenic bacteria.
Fluorescent homogeneous immunosensors for detecting pathogenic bacteria.
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DOI：
10.1016/j.ab.2009.09.039
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发表时间：
2010-01-15
影响因子：
2.9
通讯作者：
Heyduk T
中科院分区：
文献类型：
--
作者：
Heyduk E;Heyduk T
我们开发了一种简单的基于抗体的快速均相检测细菌的方法。我们的传感器利用识别靶细胞的细胞表面表位的抗体。制备两个抗体样品，每个样品通过具有短互补寡核苷酸的纳米尺寸柔性接头标记，所述短互补寡核苷酸用可参与荧光共振能量转移（FRET）的荧光染料修饰。设计互补寡核苷酸序列的长度，使得在不存在靶细胞的情况下发生非常少的退火。在靶细胞的存在下，两种标记的抗体结合到细胞表面，导致与抗体连接的互补寡核苷酸的大的局部浓度。这又驱动互补寡核苷酸的退火，其使荧光探针紧密接近，产生与靶细胞的量成比例的大FRET信号。用于将寡核苷酸连接到抗体上的长柔性连接体能够实现靶诱导的寡核苷酸退火，即使表面抗原的密度很小。我们使用了E.大肠杆菌0157：H7和鼠伤寒沙门氏菌，以证明这种设计产生的传感器表现出快速的响应时间，高特异性和检测目标细菌的灵敏度。
We developed a straightforward antibody-based assay for rapid homogeneous detection of bacteria. Our sensors utilize an antibody recognizing cell-surface epitopes of the target cell. Two samples of the antibody are prepared each labeled via nanometer size flexible linkers with short complementary oligonucleotides that are modified with fluorochromes that could participate in Fluorescence Resonance Energy Transfer (FRET). The length of the complementary oligonucleotide sequences was designed such that very little annealing occurred in the absence of the target cells. In the presence of the target cells the two labeled antibodies bind to the surface of the cell resulting in a large local concentration of the complementary oligonucleotides that are attached to the antibody. This in turn drives the annealing of the complementary oligonucleotides which brings the fluorescence probes to close proximity producing large FRET signal proportional to the amount of target cells. Long flexible linkers used to attach the oligonucleotides to the antibody enable target-induced oligonucleotide annealing even if the density of surface antigens is only modest. We used E. coli 0157:H7 and Salmonella typhimurium to demonstrate that this design produced sensors exhibiting rapid response time, high specificity and sensitivity of detecting the target bacteria.
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影响因子：
4.4
作者：
BEJ, AK;MAHBUBANI, MH;ATLAS, RM
通讯作者：
ATLAS, RM
影响因子：
2.2
作者：
Blais, BW;Bosley, J;Popela, M
通讯作者：
Popela, M
影响因子：
3.3
作者：
MAHBUBANI, MH;BEJ, AK;ATLAS, RM
通讯作者：
ATLAS, RM
影响因子：
7.4
作者：
Heyduk, E;Heyduk, T
通讯作者：
Heyduk, T
影响因子：
2.9
作者：
Tian, Ling;Heyduk, Tomasz
通讯作者：
Heyduk, Tomasz